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And Pelargonium Sidoides (DC) (Geraniaceae) Root Extracts

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And Pelargonium Sidoides (DC) (Geraniaceae) Root Extracts South African Journal of Botany 72 (2006) 232 – 237 www.elsevier.com/locate/sajb Antibacterial, antifungal and antitubercular activity of (the roots of) Pelargonium reniforme (CURT) and Pelargonium sidoides (DC) (Geraniaceae) root extracts S.P.N. Mativandlela, N. Lall *, J.J.M. Meyer Department of Botany, University of Pretoria, Pretoria 0002, South Africa Received 10 May 2005; accepted 26 August 2005 Abstract Root extracts of Pelargonium reniforme CURT and Pelargonium sidoides DC were evaluated for antibacterial and antifungal assays using the agar dilution while antitubercular assays were done using the BACTEC method at concentrations ranging from 5Â103 to 500.0 mg/L. The ethanol and acetone extracts of the roots of P. sidoides inhibited the growth of Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumoniae at a concentration of 5Â103 mg/L. Both acetone and ethanol extracts of P. reniforme and only the ethanol extract of P. sidoides inhibited the growth of Aspergillus niger and Fusarium oxysporum significantly at a concentration of 5Â103 mg/L. Growth of Rhizopus stolonifer was suppressed by the ethanol extract of P. reniforme and P. sidoides at 5Â103 and 1Â103 mg/L, respectively. Acetone, chloroform and ethanol extracts of P. reniforme showed activity against M. tuberculosis exhibiting a minimum inhibitory concentration of 5Â103 mg/L. D 2006 SAAB. Published by Elsevier B.V. All rights reserved. Keywords: Antibacterial; Antifungal; Extracts; Mycobacterium tuberculosis; Pelargonium 1. Introduction Africa use these species to treat coughs, diarrhoea and tuberculosis (Watt and Breyer-Brandwijk, 1962). The medic- The importance of Pelargonium species (Geraniaceae) is inally active ingredients are found in the bitter tasting roots of well documented (Watt and Breyer-Brandwijk, 1962; Hutch- the plants (Helmstadter, 1996). A commonly used medicine ings, 1996). The genus Pelargonium comprises more than 250 produced in Germany, named, FUmckaloabo_ originates from natural species of perennial small shrubs, which are limited in the roots of P. sidoides and P. reniforme (Helmstadter, 1996; their geographical distribution. About 80% of Pelargonium Kayser et al., 1998). This herbal medicine is extensively used species are confined to the southern parts of Africa, while in Germany for bronchitis, antibacterial and antifungal infec- others occur in Australia, New Zealand and the Far East. These tions. Although this herbal medicine (UmckaloaboR)is species usually grow in short grassland and sometimes with successfully employed in modern phytotherapy in Europe to shrubs and trees on stony soil varying from sand to clay-loam, cure infectious diseases of the respiratory tract, the scientific shale or basalt. The plants are evergreen when cultivated, but basis of its remedial effect is still unclear (Kayser and die back in nature during droughts and winter (May to August) Kolodziej, 1995). (Van der Walt and Vorster, 1985). Bacteria, which are associated with either primary or Pelargonium reniforme CURT and Pelargonium sidoides secondary infections of bronchitis, are Streptococcus pneumo- DC are highly valued by traditional healers for their curative niae, Haemophilus influenzae and Moraxella catarrhalis. H. properties and they are well known to generations of Khoi/San influenzae, a Gram-negative bacterium, is an obligate human and Xhosa (South African tribes) traditional healers (Wagner parasite that is passed from person to person by way of the and Bladt, 1975). The Xhosa and the Zulu tribes of South respiratory route. M. catarrhalis, a Gram-negative bacterium, causes bronchitis and pneumonia in children and adults. S. * Corresponding author. pneumoniae, a Gram-positive bacterium, infects the upper E-mail address: [email protected] (N. Lall). respiratory tract and can cause pneumonia, also it can infect the 0254-6299/$ - see front matter D 2006 SAAB. Published by Elsevier B.V. All rights reserved. doi:10.1016/j.sajb.2005.08.002 S.P.N. Mativandlela et al. / South African Journal of Botany 72 (2006) 232–237 233 lining of the brain-spinal cord (meningitis), bones (osteomy- respectively. Seider and Taylor (2004) investigated the two elitis), joints (arthritis), ears (otitis media) and sinuses (sinusitis plant species against rapidly growing mycobacteria (M. and bronchitis) (Benjamin et al., 1991). aurum and M. fortuitum, M. phlei, M. abscessus and M. Apergillus niger, Fusarium oxysporum and Rhizopus smegmatis). This is the first report on antitubercular activity stolonifer are some of the fungal pathogens that can affect of these plants extracted using various solvents such as the respiratory tract. A. niger, is a causative agent of pulmonary chloroform, acetone and ethanol against M. tuberculosis using diseases including aspergillosis, bronchial asthma and acute BACTEC radiometric method. allergic alveolitis. The fungus colonizes old tuberculosis or bronchiostatic cavities, in which it forms a large colony 2. Materials and methods (aspergilloma); or it may actually invades the lung tissue to produce haemorrhagic and necrotizing pneumonia (MacSween 2.1. Plant material and Whaley, 1992). F. oxysporum is responsible for fusariosis, skin infection, respiratory tract infections (tuberculosis and Roots of P. reniforme and P. sidoides were collected from bronchitis) and arthritis and produces a 76% mortality rate in Qwaqwa, a region in the Free State province of South Africa. hospitalised immunocompromised patients (Monier et al., Voucher specimens of P. reniforme (P 092558) and P. sidoides 1994). R. stolonifer causes mucorosis disease and it has been (P 092559) were deposited and identified at the H.G.W.J. reported that exposure to large numbers of Rhizopus spores can Schweickerdt Herbarium (PRU), University of Pretoria, South cause respiratory complications (Alexopoulos et al., 1996). Africa. Previously, researchers have reported antimicrobial activity of extracts of Pelargoniums and their constituents against a few 2.2. Preparation of extracts bacterial (Staphylococcus aureus, S. pneumoniae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas Air-dried and powdered roots of P. reniforme and P. aeruginosa and H. influenzae), and fungal (Microsporum sidoides (300 g each) were extracted three times with 1 L of canis, M. gypseum, A. fumigatus, Mucor racemosus, R. acetone, chloroform and ethanol separately for 2 h at room nigricans) pathogens as well as opportunistic yeasts such as temperature. The extracts were filtered through Whatman No. Candida albicans, C. glabrata, C. krusei and Cryptococcus 1 filter paper and concentrated with a rotary vacuum neoformans (Kolodziej, 2000; Kolodziej et al., 2003; Latte´ and evaporator (Bu¨chi Laboratoriums, Technik AG, Germany) to Kolodziej, 2000). Plant extracts of P. reniforme and P. sidoides dryness at reduced pressure. For antibacterial and antifungal have not been tested against the fungal pathogens, A. niger, F. assays, acetone and ethanol extracts were dissolved in oxysporum, R. stolonifer and the Gram-negative bacteria M. acetone to a concentration of 5Â104 and 1Â105 mg/L, catarrhalis, which are indirectly responsible for secondary respectively. For the antitubercular assay, all (3) extracts were infections in cases of bronchitis and tuberculosis. In the present dissolved in dimethyl sulphoxide (DMSO) to a concentration study, we have investigated their antimicrobial activity against of 5Â105 mg/L. the bacteria and fungi mainly responsible for bronchitis. We have also confirmed the findings of other researchers on the 2.3. Microorganisms and in vitro antimicrobial assays antibacterial activity of these species against S. pneumoniae and H. influenzae. 2.3.1. Bacteria Tuberculosis (TB) kills approximately 2 million people The bacteria used in this investigation H. influenzae (UPM each year, the global epidemic is growing and becoming more 2), M. catarrhalis (UPM 4), and S. pneumoniae (UPM 9) were problematic. The breakdown in health services, the spread of clinical isolates which were obtained from the Department of HIV/AIDS and the emergence of multidrug-resistant strains of Pathology, University of Pretoria, South Africa. Cultures were Mycobacterium tuberculosis (MDR) TB are contributing to maintained on Colombia agar (Oxoid, Basingstoke, UK) slants the worsening impact of this disease. It is estimated that supplemented with 5% horse blood to form chocolate agar. For between 2002 and 2020, approximately a billion people will assays, organisms were subcultured once and incubated at be newly infected, more than 150 million people will get sick, 37 -C on Mueller-Hinton (MH), (BIOLAB, Merck, South and 36 million will die of TB. The current threat in TB Africa) agar for 24 h. treatment lies in the emergence of strains resistant to two of the best antitubercular drugs, isoniazid (INH) and rifampicin 2.3.2. Antibacterial assay (RIF). The current TB-treatment comprises of 3–4 drugs for a For the antibacterial assay, the minimum inhibitory con- period of 6–9 months (Bloom, 2002). Novel drugs are centrations (MIC which is defined as the lowest concentration required which can shorten this long-treatment period and of the extract that inhibits more than 99% of the bacterial target multidrug resistant strains of TB. Previous studies have population) of the acetone and ethanol extracts were deter- investigated the anti-TB and antimycobacterial activity of the mined by incorporating various amounts (5Â103,1Â103 and two Pelargonium species. Kolodziej (2000) and Kolodziej et
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